For organs to survive storage, cell structure and function must be maintained during the ischemic period. Knowledge of the molecular events which occur during an interruption in tissue oxygen or blood supply is essential if improved techniques are to be developed. The objectives of this proposed study are to investigate the changes in nucleotide levels which accompany organ storage, and the effect of this storage on the structure and function of chromatin. A correlation has been proposed between organ function after storage and the adenine nucleotide levels. We will use high pressure liquid chromatography to examine the effects of storage on all nucleotide levels and on some essential aspects of nucleotide metabolism. A decrease in nucleotide levels would affect many levels of cell function including transcription of cell chromatin. Recent advances in knowledge of the biochemistry of chromatin have led investigators to propose a "beads on a string" structure for chromatin. Using new methods for isolation of nucleosomes (the "beads" in the chromatin structure), we will examine the effect of storage on the molecular structure of the nucleosome units and upon their ability to act as template for RNA polymerase.